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Engineering light-inducible nuclear localization signals for precise spatiotemporal control of protein dynamics in living cells

Author

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  • Dominik Niopek

    (German Cancer Research Center (DKFZ), Im Neuenheimer Feld 580
    Synthetic Biology Group, Institute for Pharmacy and Molecular Biotechnology (IPMB) and BioQuant, University of Heidelberg, Im Neuenheimer Feld 267)

  • Dirk Benzinger

    (Synthetic Biology Group, Institute for Pharmacy and Molecular Biotechnology (IPMB) and BioQuant, University of Heidelberg, Im Neuenheimer Feld 267
    Present address: Department for Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland)

  • Julia Roensch

    (German Cancer Research Center (DKFZ), Im Neuenheimer Feld 580)

  • Thomas Draebing

    (Synthetic Biology Group, Institute for Pharmacy and Molecular Biotechnology (IPMB) and BioQuant, University of Heidelberg, Im Neuenheimer Feld 267
    Present address: Department of Internal Medicine III, University Clinic Heidelberg, Im Neuenheimer Feld 669, 69120 Heidelberg, Germany)

  • Pierre Wehler

    (German Cancer Research Center (DKFZ), Im Neuenheimer Feld 580)

  • Roland Eils

    (German Cancer Research Center (DKFZ), Im Neuenheimer Feld 580
    Synthetic Biology Group, Institute for Pharmacy and Molecular Biotechnology (IPMB) and BioQuant, University of Heidelberg, Im Neuenheimer Feld 267)

  • Barbara Di Ventura

    (Synthetic Biology Group, Institute for Pharmacy and Molecular Biotechnology (IPMB) and BioQuant, University of Heidelberg, Im Neuenheimer Feld 267)

Abstract

The function of many eukaryotic proteins is regulated by highly dynamic changes in their nucleocytoplasmic distribution. The ability to precisely and reversibly control nuclear translocation would, therefore, allow dissecting and engineering cellular networks. Here we develop a genetically encoded, light-inducible nuclear localization signal (LINuS) based on the LOV2 domain of Avena sativa phototropin 1. LINuS is a small, versatile tag, customizable for different proteins and cell types. LINuS-mediated nuclear import is fast and reversible, and can be tuned at different levels, for instance, by introducing mutations that alter AsLOV2 domain photo-caging properties or by selecting nuclear localization signals (NLSs) of various strengths. We demonstrate the utility of LINuS in mammalian cells by controlling gene expression and entry into mitosis with blue light.

Suggested Citation

  • Dominik Niopek & Dirk Benzinger & Julia Roensch & Thomas Draebing & Pierre Wehler & Roland Eils & Barbara Di Ventura, 2014. "Engineering light-inducible nuclear localization signals for precise spatiotemporal control of protein dynamics in living cells," Nature Communications, Nature, vol. 5(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms5404
    DOI: 10.1038/ncomms5404
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    Cited by:

    1. Yage Ding & Cristina Tous & Jaehoon Choi & Jingyao Chen & Wilson W. Wong, 2024. "Orthogonal inducible control of Cas13 circuits enables programmable RNA regulation in mammalian cells," Nature Communications, Nature, vol. 15(1), pages 1-16, December.

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